We plan to continue our investigation on oxidative metabolism in guinea pig and human granulocytes. We will make rats either selenium or riboflavin deficient and examine the effects of these deficiencies on granulocyte O2\negative production, hexosemonophosphate shunt activity and plasma membrane NAD(P)H oxidase in addition to other aspects of phagocytosis. We plan to improve our isolation techniques for plasma membranes and examine the submembranous location for the O2\negative-generating NAD(P)H oxidase. We also plan to characterize this enzyme that we find in phagocytic vesicles. Since the oxidase is normally inactivated after 7-10 minutes of activity, we plan to examine the effect of oxygen and its metabolites on this inactivation. We also will attempt to label this enzyme with a radioactive photoaffinity probe. This will allow us to learn more about this enzyme as a protein and may aid in our ability to isolate the enzyme and identify it in granulocytes from patients with chronic granulomatous disease.